Abstract：
BACKGROUND: While 15 to 20% of cancers are associated with microbial infection, the relationship between oral microorganisms and oral squamous cell carcinoma (OSCC) remains unclear. The location of bacteria in a tumor is closely related to its carcinogenic mechanism. The aim of this study was to analyse bacterial diversity in clinical OSCC tissue samples and tumor distant normal tissues, locate target bacteria, and search for proteins that may interact with target bacteria.
METHODS: The 16S rDNA method was used to analyse bacterial diversity in clinical OSCC tissue samples and tumor distant normal tissues. Correlations between Fusobacterium abundance and clinicopathological characteristics were analysed using the χ2 test. The position of target bacteria was analysed by fluorescence in situ hybridization (FISH), and the expression of CK, CD31, CD45, CD68, cyclin D1, β-catenin, E-cadherin, NF-κB, and HIF-1α was analysed by immunohistochemistry (IHC) in OSCC tumor tissues and tumor distant normal tissues.
RESULTS: The 16S rDNA results showed that the detected amount of Fusobacterium in OSCC tumor tissues was significantly larger than that in tumor distant normal tissues. High expression of Fusobacterium was significantly correlated with the lifestyle-related oral risk habits, including smoking (p=0.036) and alcohol consumption (p=0.022), but did not correlate with patient sex, age, tumor laterality, tumor size, grade or TNM stage. Fusobacterium nucleatum was enriched in tumor stroma, where CD31+ blood vessels and inflammatory cells (including CD45+ leukocytes and CD68+ macrophages) were densely distributed. Cyclin D1 was mainly expressed in the nucleus of tumor cells. β-catenin was expressed in the tumor cell membrane and was positively expressed in tumor interstitial vascular endothelial cells. E-cadherin was mainly expressed in tumor cell membranes. NF-κB was positively expressed in the cytoplasm of tumor cells, tumor interstitial cells and myo-fibrocytes. HIF-1α was mainly expressed in the cytoplasm of tumor interstitial cells. HIF-1α was highly expressed where Fusobacterium nucleatum was densely distributed.
CONCLUSIONS: According to our study, the detected amount of Fusobacterium in OSCC tumor tissues was significantly larger than that in tumor distant normal tissues, and Fusobacterium nucleatum might aggravate inflammation and hypoxia by interacting with NF-κB and HIF-1α in OSCC.
METHODS: The 16S rDNA method was used to analyse bacterial diversity in clinical OSCC tissue samples and tumor distant normal tissues. Correlations between Fusobacterium abundance and clinicopathological characteristics were analysed using the χ2 test. The position of target bacteria was analysed by fluorescence in situ hybridization (FISH), and the expression of CK, CD31, CD45, CD68, cyclin D1, β-catenin, E-cadherin, NF-κB, and HIF-1α was analysed by immunohistochemistry (IHC) in OSCC tumor tissues and tumor distant normal tissues.
RESULTS: The 16S rDNA results showed that the detected amount of Fusobacterium in OSCC tumor tissues was significantly larger than that in tumor distant normal tissues. High expression of Fusobacterium was significantly correlated with the lifestyle-related oral risk habits, including smoking (p=0.036) and alcohol consumption (p=0.022), but did not correlate with patient sex, age, tumor laterality, tumor size, grade or TNM stage. Fusobacterium nucleatum was enriched in tumor stroma, where CD31+ blood vessels and inflammatory cells (including CD45+ leukocytes and CD68+ macrophages) were densely distributed. Cyclin D1 was mainly expressed in the nucleus of tumor cells. β-catenin was expressed in the tumor cell membrane and was positively expressed in tumor interstitial vascular endothelial cells. E-cadherin was mainly expressed in tumor cell membranes. NF-κB was positively expressed in the cytoplasm of tumor cells, tumor interstitial cells and myo-fibrocytes. HIF-1α was mainly expressed in the cytoplasm of tumor interstitial cells. HIF-1α was highly expressed where Fusobacterium nucleatum was densely distributed.
CONCLUSIONS: According to our study, the detected amount of Fusobacterium in OSCC tumor tissues was significantly larger than that in tumor distant normal tissues, and Fusobacterium nucleatum might aggravate inflammation and hypoxia by interacting with NF-κB and HIF-1α in OSCC.
摘要：
背景：虽然15％至20％的癌症与微生物感染有关，口腔微生物与口腔鳞状细胞癌（OSCC）之间的关系尚不清楚。细菌在肿瘤中的位置与其致癌机制密切相关。这项研究的目的是分析临床OSCC组织样本和肿瘤远处正常组织中的细菌多样性，定位目标细菌，寻找可能与目标细菌相互作用的蛋白质。
方法：使用16SrDNA方法分析临床OSCC组织样本和肿瘤远处正常组织中的细菌多样性。使用χ2检验分析了梭杆菌丰度与临床病理特征之间的相关性。通过荧光原位杂交（FISH）分析目标细菌的位置，和CK的表达，CD31，CD45，CD68，细胞周期蛋白D1，β-连环蛋白，E-cadherin,NF-κB,免疫组织化学（IHC）分析OSCC肿瘤组织和肿瘤远处正常组织中的HIF-1α。
结果：16SrDNA结果显示，OSCC肿瘤组织中梭杆菌属的检出量明显大于肿瘤远端正常组织。梭菌的高表达与生活方式相关的口腔风险习惯显着相关，包括吸烟（p=0.036）和饮酒（p=0.022），但与患者性别无关，年龄,肿瘤侧向性，肿瘤大小,等级或TNM阶段。核梭杆菌在肿瘤基质中富集，其中CD31+血管和炎性细胞(包括CD45+白细胞和CD68+巨噬细胞)密集分布。CyclinD1主要在肿瘤细胞核中表达。β-catenin在肿瘤细胞膜上表达，在肿瘤间质血管内皮细胞中呈阳性表达。E-cadherin主要在肿瘤细胞膜上表达。NF-κB在肿瘤细胞胞浆中呈阳性表达,肿瘤间质细胞和肌纤维细胞。HIF-1α主要表达于肿瘤间质细胞的胞浆中。HIF-1α在核梭杆菌密集分布的地方高表达。
结论：根据我们的研究，OSCC肿瘤组织中梭杆菌的检出量明显大于肿瘤远处正常组织,核梭杆菌可能通过与NF-κB和HIF-1α相互作用而加重OSCC的炎症和缺氧。
方法：使用16SrDNA方法分析临床OSCC组织样本和肿瘤远处正常组织中的细菌多样性。使用χ2检验分析了梭杆菌丰度与临床病理特征之间的相关性。通过荧光原位杂交（FISH）分析目标细菌的位置，和CK的表达，CD31，CD45，CD68，细胞周期蛋白D1，β-连环蛋白，E-cadherin,NF-κB,免疫组织化学（IHC）分析OSCC肿瘤组织和肿瘤远处正常组织中的HIF-1α。
结果：16SrDNA结果显示，OSCC肿瘤组织中梭杆菌属的检出量明显大于肿瘤远端正常组织。梭菌的高表达与生活方式相关的口腔风险习惯显着相关，包括吸烟（p=0.036）和饮酒（p=0.022），但与患者性别无关，年龄,肿瘤侧向性，肿瘤大小,等级或TNM阶段。核梭杆菌在肿瘤基质中富集，其中CD31+血管和炎性细胞(包括CD45+白细胞和CD68+巨噬细胞)密集分布。CyclinD1主要在肿瘤细胞核中表达。β-catenin在肿瘤细胞膜上表达，在肿瘤间质血管内皮细胞中呈阳性表达。E-cadherin主要在肿瘤细胞膜上表达。NF-κB在肿瘤细胞胞浆中呈阳性表达,肿瘤间质细胞和肌纤维细胞。HIF-1α主要表达于肿瘤间质细胞的胞浆中。HIF-1α在核梭杆菌密集分布的地方高表达。
结论：根据我们的研究，OSCC肿瘤组织中梭杆菌的检出量明显大于肿瘤远处正常组织,核梭杆菌可能通过与NF-κB和HIF-1α相互作用而加重OSCC的炎症和缺氧。
